Oligonucleotides For the Detection of Hepatitis B Virus

• US 20080280287A1
• Filed: 12/07/2004
• Published: 11/13/2008
• Est. Priority Date: 12/16/2003
• Status: Abandoned Application

• Current Assignee: Bio-Rad Innovations.
• Original Assignee: Bio-Rad Innovations.
• Inventors: Biron, Marie-Philippe
• Application Number:
  US10/583,088
• Publication Number:
  US 20080280287A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/701 Specific hybridization probes
• CPC Class Codes: C12Q 1/706 for hepatitis
• CPC Class Codes: C12Q 2531/113 PCR
• CPC Class Codes: C12Q 2561/101 Taqman
• CPC Class Codes: C12Q 2561/113 Real time assay

Abstract

The invention relates to specific oligonucleotide primers and probes for rapid and sensitive detection of hepatitis B virus by amplification of the HBV nucleic acid, and to methods using the same. It is also concerned with kits useful to perform these detection tests.

Claims

1. :

An oligonucleotide that includes a sequence selected from the group consisting of SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7 and their complementary sequences.

2. :

The oligonucleotide according to claim 1, which consists of SEQ ID NO;

2 or its complementary sequence.

3. :

The oligonucleotide according to claim 1, which consists of SEQ ID NO;

3, or its complementary sequence.

4. :

The oligonucleotide according to claim 1, which consists of SEQ ID NO;

4, or its complementary sequence.

5. :

The oligonucleotide according to claim 1, which consists of SEQ ID NO;

5, or its complementary sequence.

6. :

The oligonucleotide according to claim 1, which consists of SEQ ID NO;

6, or its complementary sequence.

7. :

The oligonucleotide according to claim 1, which consists of SEQ ID NO;

7, or its complementary sequence.

8. :

Use of an oligonucleotide as defined in claim 1, as a probe or primer, for hybridizing with and optionally amplifying a nucleic acid from a hepatitis B virus (HBV).

9. :

Use of an oligonucleotide that includes a sequence selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, and their complementary sequence, as a probe for hybridizing with a nucleic acid from HBV.

10. :

The use according to claim 9, wherein said oligonucleotide consists of a sequence selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, and their complementary sequence.

11. :

The use according to claim 9, wherein said oligonucleotide includes a sequence selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, and their complementary sequence and carries a fluorophore moiety at one terminus, and a quencher moiety at the other terminus.

12. :

The use according to claim 11, wherein said oligonucleotide consists of a sequence selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14 and SEQ ID NO;

15, and carries a fluorophore moiety at one terminus, and a quencher moiety at the other terminus.

13. :

A set of oligonucleotides consisting of an oligonucleotide that includes SEQ ID NO;

2, and at least an oligonucleotide selected from the group consisting of an oligonucleotide that includes SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6 and SEQ ID NO;

7.

14. :

A set of oligonucleotides according to claim 13, which consists of;

(i) an oligonucleotide that includes SEQ ID NO;

2, and an oligonucleotide that includes SEQ ID NO;

3;

(ii) an oligonucleotide that includes SEQ ID NO;

2, and an oligonucleotide that includes SEQ ID NO;

4;

(iii) an oligonucleotide that includes SEQ ID NO;

2, and an oligonucleotide that includes SEQ ID NO;

5;

(iv) an oligonucleotide that includes SEQ ID NO;

2, and an oligonucleotide that includes SEQ ID NO;

6;

(v) an oligonucleotide that includes SEQ ID NO;

2, and an oligonucleotide that includes SEQ ID NO;

7;

(vi) an oligonucleotide that includes SEQ ID NO;

2, an oligonucleotide that includes SEQ ID NO;

4 and an oligonucleotide that includes SEQ ID NO;

5; and

(vii) an oligonucleotide that includes SEQ ID NO;

2, an oligonucleotide that includes SEQ ID NO;

6 and an oligonucleotide that includes SEQ ID NO;

7.

15. :

A set of oligonucleotides comprising;

a) a set of oligonucleotides according to claim 13; and

b) an oligonucleotide that includes a sequence selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, and their complementary sequence.

16. :

A set of oligonucleotides that comprises;

a) a set of oligonucleotides according to claim 13; and

b) an oligonucleotide that consists of a sequence selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14 and SEQ ID NO;

15, and carries a fluorophore moiety at one terminus, and a quencher moiety at the other terminus.

17. :

A method for specifically detecting a HBV by amplification in a biological sample, which method comprises the steps consisting of;

a) contacting a set of oligonucleotides according to claim 13 with a biological sample or nucleic acid preparation obtained from a biological sample, under conditions suitable for the oligonucleotides to hybridize to a HBV nucleic acid present in the sample;

b) amplifying said HBV nucleic acid using said oligonucleotides as primers;

c) detecting the amplification product, indicative of the presence of a HBV in the biological sample.

18. :

The method according to claim 17, wherein HBV nucleic acid is amplified by polymerase chain reaction.

19. :

The method according to claim 17, wherein the detection of said amplification product is performed by using an oligonucleotide that includes a sequence selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, and their complementary sequence, and that is detectably labelled, as a probe.

20. :

The method according to claim 19, wherein said oligonucleotide that includes a sequence selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, and their complementary sequence, and carries a fluorophore moiety at one terminus, and a quencher moiety at the other terminus.

21. :

The method according to claim 19, wherein said oligonucleotide that includes a sequence selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, and their complementary sequence, is SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14 or SEQ ID NO;

15.

22. :

A kit for amplifying HBV in a biological sample, which kit comprises;

at least a set of oligonucleotides according to claim 13, useful as primers;

means for amplifying a HBV nucleic acid.

23. :

The kit according to claim 22, which further comprises means for the detection of the amplified product.

24. :

The kit according to claims 22, wherein the means for amplifying HBV nucleic acid are means for amplification by Polymerase Chain Reaction.

25. :

The kit according to claim 22, which comprises an oligonucleotide that includes a sequence selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, and their complementary sequence, detectably labelled and useful as a probe.

26. :

The kit according to claim 25, wherein said oligonucleotide that includes a sequence selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, and their complementary sequence, is SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14 or SEQ ID NO;

15.